Effect of different matrices on probiotic resistance to in vitro simulated gastrointestinal conditions

In this study, we evaluated the effect of different matrices (MRS, milk and milk with inulin) on the tolerance of probiotic strains (Lactobacillus acidophilus La‐5 and Bifidobacterium animalis subsp. lactis BB‐12) to simulated conditions similar to those found in the gastrointestinal tract. Both probiotic strains demonstrated a significantly lower viability after exposure to in vitro gastric and intestinal conditions, and B. animalis subsp. lactis BB‐12 showed higher survival during the test compared to L. acidophilus La‐5 in all tested matrices. Milk and inulin protected probiotics from in vitro gastrointestinal stress. These results suggest that it is critical to formulate the food matrix to be used as probiotic carrier.     

Lactic acid starter and probiotic bacteria: a comparative “in vitro” study of probiotic characteristics and biological barrier resistance

Probiotic characteristics (deconjugation of bile salts, hydrophobicity and β-galactosidase activity) and the resistance to biological barriers (gastric juice and bile salts) of 24 strains of lactic acid starter bacteria (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and Lactococcus lactis) and 24 strains of probiotic bacteria (Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus and bifidobacteria) were compared. Among the probiotic bacteria tested, Lactobacillus acidophilus was the most interesting species since it showed high values of resistance to gastric juice and bile, hydrophobicity and β-galactosidase and bile salts deconjugation activities. Bifidobacterium bifidum strains showed the same behavior, although the values of the parameters investigated were slightly lower than those obtained for Lactobacillus acidophilus. On the other hand, it was demonstrated that Lactobacillus delbrueckii subsp. bulgaricus was the lactic acid starter species with the best probiotic characteristics among the starter species assessed. It was resistant to gastric juice and bile, and showed high values for β-galactosidase activity. On the other hand, lactic acid starter bacteria showed hydrophobicity values similar to or higher than those obtained for the strains of the Lactobacillus casei . According to the results found, the total probiotic value of a fermented dairy product should take into account not only the intestinal probiotic cultures used in the formulation but also the probiotic contribution of the lactic acid starter microflora.     

In vitro comparison of probiotic properties of Lactobacillus casei Zhang, a potential new probiotic, with selected probiotic strains

Since 2004, our research group has isolated 240 Lactobacillus strains from Koumiss, a traditional fermented alcoholic beverage prepared from mare's milk in Inner Mongolia, Xinjiang of China and Mongolia. Among these Lactobacillus strains a novel strain with potential probiotic properties, Lactobacillus casei Zhang, was screened out and studied extensively for its probiotic properties, health-promoting effects and fermentation characteristics. In vitro tests indicated that L. casei Zhang had high tolerance to simulated gastric, intestine juices and bile salts, similar to commercial probiotic strains such as Lactobacillus acidophilus NCFM, Lactobacillus rhamnosus GG, L. casei Shirota and Bifidobacterium animalis Bb12. Higher acid-production activity and proteolytic activity was observed in the fermented milk inoculated with L. casei Zhang during the refrigerated storage than in the samples inoculated with the selected commercial probiotics. The yogurt samples fermented with L. casei Zhang exhibited similarer viable count (1.0 × 10⁹ cfu/mL) as the other samples after 28 d of refrigerated storage. The results suggested that L. casei Zhang showed good potential for application in functional foods and health-related products.     

The viability of three probiotic organisms grown with yoghurt starter cultures during storage for 21 days at 4°C

This study investigated the viability of probiotic ( Lactobacillus acidophilus LA5, Lactobacillus rhamnosus LBA and Bifidobacterium animalis subsp . lactis BL-04) in milk fermented with Lactobacillus delbrueckii subsp . bulgaricus LB340 and Streptococcus thermophilus TAO (yoghurt – Y). Each probiotic strain was grown separately in co-culture with Y and in blends of different combinations. Blends affected fermentation time(s), pH and firmness during storage at 4°C. The product made with Y plus B. animalis subsp . lactis and L. rhamnosus had counts of viable cells at the end of shelf life that met the minimum required to achieve probiotic effect. However, L. acidophilus and L. delbrueckii subsp . bulgaricus were inhibited.     

Probiotic properties of novel probiotic Levilactobacillus brevis KU15147 isolated from radish kimchi and its antioxidant and immune-enhancing activities

This study was conducted to evaluate the probiotic properties and antioxidant activities of lactic acid bacteria strains including Levilactobacillus brevis KU15147 isolated from kimchi to determine their potential as a probiotic. The tolerance of all strains to gastric acid and bile salts was more than 90%. The strains did not produce a β-glucuronidase and survived following treatment with gentamicin, kanamycin, streptomycin, and ciprofloxacin. L. brevis KU15147 showed greater adhesion activity to HT-29 cells (6.38%) and its antioxidant activities were higher than those of other tested strains, showing values of 38.56%, 22%, and 23.82% in DPPH, ABTS, and β-carotene bleaching assays, respectively. Additionally, the relative expression intensities of induced nitric oxide synthase and tumor necrosis factor-α of L. brevis KU15147 were greater than those of other strains, suggesting that this strain can be applied in the health food or pharmaceutical industry as a novel probiotic strain.     

Assessment of the probiotic potential of lactic acid bacteria isolated from Minas artisanal cheese produced in the Campo das Vertentes region,Brazil

The microbiota of the Brazilian Minas artisanal cheese, made from raw milk, is not well known and may include probiotic bacteria. This study aimed to assess the in vitro and in vivo probiotic properties of the lactic acid bacteria isolated from these cheeses. Thirty‐six samples of the Lactobacillus/Pediococcus group were selected for in vitro investigation. Pediococcus acidilactici (PA2) and Lactobacillus plantarum (LP4) showed the best results and were tested for their ability to protect Salmonella Typhimurium orally infected mice. LP4 showed better probiotic potential than PA2 and allowed higher values of weight gain (P < 0.05). Thus, Lb. plantarumLP4 may have potential use as a probiotic in foods after future technological screening.     

Pediococcus acidilactici as a potential probiotic to be used in food industry

The objective of this study was to compare the results obtained between a commercial probiotic bacterium and a strain of Pediococcus acidilactici isolated from a food matrix. Besides the characterisation of the isolates, the main focus of this work was to evaluate their ability to survive through simulated gastrointestinal tract passage as well as the effect of the spray drying process to be used as concentrated cultures in the food industry. Both lactic acid bacteria produced any of the virulence determinants investigated; they exhibited no significant antibiotic resistances and <1 log unit reduction in viable counts was obtained after exposure to simulated gastrointestinal tract conditions. Concentrated cultures of P. acidilactici HA‐6111‐2 in 10% (w/v) of reconstituted skim milk were obtained by spray drying and, after 60 days of storage at 4 °C, <1 log unit reduction in viable count was recorded. In conclusion, from these preliminary characterisation tests and its extended viability in the dried form, this P. acidilactici strain should be considered as a potentially useful probiotic.     

Bioutilisation of whey for lactic acid production

The disposal of whey, the liquid remaining after the separation of milk fat and casein from whole milk, is a major problem for the dairy industry, which demands simple and economical solutions. The bioconversion of lactose present in whey to valuable products has been actively explored. Since whey and whey permeates contain significant quantities of lactose, an interesting way to upgrade this effluent could be as a substrate for fermentation. Production of lactic acid through lactic acid bacteria could be a processing route for whey lactose and various attempts have been made in this direction. Immobilised cell technology has also been applied to whey fermentation processes, to improve the economics of the process. A fermentative means of lactic acid production has advantages over chemical synthesis, as desirable optically pure lactic acid could be produced, and the demand for optically pure lactic acid has increased considerably because of its use in the production of poly(lactic acid), a biodegradable polymer, and other industrial applications. This review focuses on the various biotechnological techniques that have used whey for the production of lactic acid.     

Screening of some potentially probiotic lactic acid bacteria for their ability to synthesis conjugated linoleic acid

Strains of potentially probiotic lactobacilli, propionibacteria, leuconostoc, lactococcus, enterococcus, and pediococcus, were tested for their ability to convert linoleic acid to conjugated linoleic acid (CLA). Growth and CLA production were followed during incubation for 48 h in reconstituted skim milk containing 0.2% lipolysed sesame oil. Lactococcus lactis subsp. lactis biovar diacetylactis and Leuconostoc mesenteroides subsp. mesenteroides gave the highest CLA production. Also, the effect of lipolysed oil concentration on the growth and CLA production of six strains were studied in medium containing 0.0–1% lipolysed oil. Leuconostoc mesenteroides subsp. mesenteroides and Lac. lactis subsp. lactis biovar diacetylactis gave maximum dienes in medium containing 0.6% and 0.8% lipolysed oil respectively.     

Comparative survival and evaluation of functional probiotic properties of spray-dried lactic acid bacteria

The effect of spray drying on the viability and retention of key probiotic properties like acid and bile tolerance and cholesterol assimilation of three probiotic lactic acid bacteria (LAB) ( Lactobacillus plantarum CFR 2191, Lactobacillus salivarius CFR 2158 and Pediococcus acidilactici CFR 2193) has been studied. More than 97% survival was exhibited by the three LAB (1% cell suspension) spray dried with maltodextrin and nonfat skimmed (NFSM) as carriers. LAB cultures spray dried with maltodextrin showed significantly greater ( P ≤  0.05) retention of functional properties than those with NFSM. The results highlight a cost-effective way of producing large quantities of selected probiotic cultures with functional properties for neutraceutical application.     

Folate‐producing lactic acid bacteria from cow’s milk with probiotic characteristics

The probiotic potential of newly isolated lactic acid bacteria from cow’s milk was analysed and the efficacy of the selected isolates to produce folic acid was determined by microbiological assay. Two isolates, CM 22 and CM 28 were selected for folate production in skim milk medium and the net folate yield was 12.5 ng and 14.2 ng/mL respectively. The two isolates (CM 22 and CM 28) showed more than 98% similarity to Lactococcus subsp. cremoris and Lactococcus lactis subsp. lactis respectively by 16S rRNA sequencing.     

Evaluation of culture media for selective enumeration of probiotic strains of lactobacilli and bifidobacteria in combination with yoghurt or cheese starters

A series of culture media were evaluated for selective enumeration of 10 commercial probiotic cultures. The media M17 and MRS 5.2 were most suitable to enumerate the yoghurt starters Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, respectively. For the enumeration of probiotic Lb. acidophilus cultures, the MRS-clindamycin medium was found to be most optimal. Selective recovery results of the tested Bifidobacterium sp. were not uniform but indicated strain-to-strain variations depending on whether NPLN or MRS-LP were used. For the enumeration of probiotic cultures of Lb. rhamnosus and Lb. paracasei, LC medium (for yoghurt products) and MRS-AC medium (for cheese products) are recommended. In conclusion, our data indicate that the choice of culture medium and methodology for selective enumeration of commercial probiotic strains in combination with starters depends strongly on the product matrix, the target group and the taxonomic diversity of the bacterial background flora in the product.     

Enhancement of survival of probiotic and non-probiotic lactic acid bacteria by yeasts in fermented milk under non-refrigerated conditions

The effects of yeasts on the survival of probiotic and non-probiotic lactic acid bacteria (LAB) were studied in fermented milk under non-refrigerated conditions (30 °C) with a view to develop ambient-stable fermented milk with live LAB. Five yeasts tested (Saccharomyces bayanus, Williopsis saturnus var. saturnus, Yarrowia lipolytica, Candida kefyr and Kluyveromyces marxianus) enhanced the survival of Lactobacillus bulgaricus (but not Streptococcus thermophilus) in a mixed yoghurt culture in yoghurt by ~ 10² to 10⁵-fold. Seven yeasts examined (Candida krusei, Geotrichum candidum, Pichia subpelliculosa, Kloeckera apiculata, Pichia membranifaciens, Schizosaccharomyces pombe and Y. lipolytica) improved the survival of Lactobacillus rhamnosus in fermented milk by ~ 10³ to 10⁶-fold. W. saturnus var. saturnus enhanced the survival of Lactobacillus acidophilus, L. rhamnosus (probiotic) and Lactobacillus reuteri by up to 10⁶-fold, but the same yeast failed to improve the survival of Lactobacillus johnsonii (probiotic), S. thermophilus and L. bulgaricus in fermented milk. These results provide definitive evidence that yeasts possess stability-enhancing effects on LAB and that the specific effects of yeasts on LAB stability vary with yeasts as well as with LAB. However, the molecular mechanism of such interaction of yeasts with LAB remains to be found.     

添加乳酸片球菌AS185的益生菌Ricotta干酪品质评价

将一株益生性乳酸片球菌（Pediococcus acidilactici）AS185添加到Ricotta干酪中,通过理化指标、质构特性、乳酸菌活菌数、挥发性成分分析等评价AS185对干酪品质的影响。结果表明,经4 ℃冷藏21 d后,添加乳酸片球菌AS185的益生菌乳清干酪的脂肪、蛋白质和水分质量分数分别为17.60%、27.47%和49.95%,pH值为4.75,均与未添加益生菌的对照干酪无显著差异。贮藏21 d,乳酸菌活菌数为7.45 log CFU/g,符合益生菌产品要求;益生菌乳清干酪的硬度、粘聚性、胶着度和咀嚼度分为723.85 N、0.97 mm、699.58和699.61 mJ,均高于对照干酪,其质地更为结实,更具有咀嚼感。在挥发性成分的种类和含量方面,益生菌乳清干酪共检测出15类成分,其中酸类、醇类、酮类的相对含量较高,分别为38.56%、24.64%、13.76%,均高于对照组;在外观色泽、滋气味和组织状方面,益生菌干酪与对照干酪无显著差异（p>0.05）,但整体可接受性方面优于对照组,说明益生菌乳酸片球菌AS185的添加对干酪风味的形成有积极作用。乳酸片球菌AS185具有适合Ricotta乳清干酪加工的技术特性,是一株有应用前景的益生菌菌株。     

Effect of bacteriocin‐producing Pediococcus acidilactici K10 on beer fermentation

Beer is generally considered to be a beverage that has high microbiological stability. However, some undesirable lactic acid bacteria (LAB) can grow in beer and consequently spoil this beverage. In this study, bacteriocin‐producing Pediococcus acidilactici K10 was used as a means of bio‐acidifying the mash and reducing the spoilage LAB content of the beer. The K10 strain had antimicrobial activity against two beer spoilage LAB strains in wort and did not grow in a beer environment. The K10 strain was inoculated before the mashing step. The effect of K10 as a starter culture was investigated and compared with a control. As a result, filtration time was shortened by 17 min, alcohol content was increased by 137%, foam stability was increased by 156%, bitterness was increased by two bitterness units and there was a significant difference (p < 0.05) in aromatic and sour odour. The feasibility of using bacteriocin‐producing LAB strain in beer brewing is envisaged. Copyright © 2016 The Institute of Brewing & Distilling     

Pediococcus acidolactici and Pediococcus pentosaceus isolated from a rainbow trout ecosystem have probiotic and ABF1 adsorbing/degrading abilities in vitro

ABSTRACT

Probiotics are being used in biological control of bacterial pathogens, as an alternative to antibiotics, to improve health and production parameters in fish farming. Fish farming production is severely affected by aflatoxins (AFs), which are a significant problem in aquaculture systems. Aflatoxins exert substantial impact on production, causing disease with high mortality and a gradual decline of reared fish stock quality. Some aspects of aflatoxicosis in fish, particularly its effects on the gastrointestinal tract, have not been well documented. The aim of the present study was to evaluate probiotic properties of lactic acid bacterial (LAB) strains isolated from rainbow trout intestine and feed. Moreover, AFB₁-binding and/or degrading abilities were also evaluated to assess their use in the formulation of feed additives. Growth at pH 2, the ability to co-aggregate with bacterial pathogens, inhibition of bacterial pathogens, and determination of the inhibitory mechanism were tested. Aflatoxin B₁ (AFB₁) adsorption and degradation ability were also tested. All strains were able to maintain viable (10⁷ cells ml^–1) at pH 2. Pediococcus acidilactici RC001 and RC008 showed the strongest antimicrobial activity, inhibiting all the pathogens tested. The strains produced antimicrobial compounds of different nature, being affected by different treatments (catalase, NaOH and heating), which indicated that they could be H₂O_2, organic acids or proteins. All LAB strains tested showed the ability to coaggregate pathogenic bacteria, showing inhibition percentages above 40%. Pediococcus acidilactici RC003 was the one with the highest adsorption capacity and all LAB strains were able to degrade AFB₁ with percentages higher than 15%, showing significant differences with respect to the control. The ability of some of the LAB strains isolated in the present work to compete with pathogens, together with stability against bile and gastric pH, reduction of bioavailability and degradation of AFB₁, may indicate the potential of LAB for use in rainbow trout culture.     

Screening of lactic acid bacteria and bifidobacteria for potential probiotic use in Iberian dry fermented sausages

The purpose of this study was to select lactic acid bacteria and bifibobacteria strains as potential probiotic cultures during the processing of Iberian dry fermented sausages. A total of 1000 strains were isolated from Iberian dry fermented sausages (363), and human (337) and pig faeces (300) in different culture media. Around 30% of these strains, mainly isolated from Iberian dry fermented sausages in LAMVAB agar, were pre-selected for testing as potential probiotics by their ability to grow adequately at the pH values and NaCl concentrations of these meat products during the ripening process. Of the in vitro investigations used to predict the survival of a strain in conditions present in the gastro intestinal tract, exposure to pH 2.5 showed itself to be a highly discriminating factor with only 51 out of 312 pre-selected strains resisting adequately after 1.5h of exposure. All acid-resistant isolates identified as lactobacilli originated from human faeces (Lactobacillus casei and Lactobacillus fermentum) and pig faeces (Lactobacillus reuteri, Lactobacillus animalis, Lactobacillus murinus, and Lactobacillus vaginalis). Pediococcus acidilactici strains were isolated from Iberian dry fermented sausages and pig faeces, whereas the greatest number of Enterococcus strains were identified as Enterococcus faecium, with this species being isolated from Iberian dry fermented sausages, and human and pig faeces. Most of these strains are promising probiotic meat culture candidates suitable for Iberian dry fermented sausages.     

Manufacture and characterization of functional emmer beverages fermented by selected lactic acid bacteria

Autochthonous lactic acid bacteria from emmer flour were screened based on the kinetic of acidification and used to ferment beverages containing emmer flour, emmer gelatinized flour, and emmer malt at percentages ranging 5-30% (wt/wt). Preliminarily, the concentration of raw flour and malt was selected based on sensory analysis. Different protocols were set up for the manufacture of four different beverages which used Lactobacillus plantarum 6E as the starter. Emmer beverages were mainly differentiated based on the concentration of organic acids, carbohydrates, amino acids, dietary fibers, vitamins, antioxidant and phytase activities, and volatiles and sensory profiles. Wheat flour bread was used as the control to determine the hydrolysis index (HI = 100), as an indirect estimation of the glycemic index. The beverage made with 30% (wt/wt) of gelatinized flour showed an HI of 56%, its viscosity was improved by using an EPS-producing strain and it allowed the survival of the potential probiotic Lactobacillus rhamnosus SP1 at cell density of ca. 5 × 10⁸ cfu/ml throughout storage at 4 °C. Among the exploited biotechnological options, this latter beverage could be considered as a promising novel functional food.     

In vitro probiotic evaluation of potential antioxidant lactic acid bacteria isolated from idli batter fermented with Piper betle leaves

Lactic acid bacteria isolated from food sources can be introduced as probiotics because of their health‐promoting and nonpathogenic characteristics. Eight lactic acid bacteria from idli batter fermented with Piper betle leaves were selected to screen the antioxidant potential and biofilm‐forming ability. Five isolates exhibited good antioxidant potential and biofilm formation and showed antagonistic activity against the pathogenic biofilm‐forming Staphylococcus aureus. Further, in vitro probiotic properties of five isolates exhibited excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, auto‐aggregation and co‐aggregation and hydrophobicity and showed resistance to antibiotics such as ciprofloxacin, nalidixic acid, norfloxacin and colistin. Isolates also showed positive bile salt hydrolase activity, cholesterol assimilation, β‐galactosidase production and mucin binding ability. The isolate KJBB10 exhibited good adhesion and pathogen exclusion percentage with human colon cancer cells (HCT‐15 and HT‐29). Hence, the isolates KJBB10, KJBB56 and KJBC06 can be used as beneficial probiotic starter cultures for the formulation of functional foods.     

乳酸菌的益生功能及作用机制研究进展

乳酸菌类益生菌是功能性食品的主要生理活性成分之一,其益生作用已经成为食品科学和临床医学领域的研究热点。本文就近年来乳酸菌改善乳糖不耐症、防治腹泻和消化性溃疡、提高机体免疫力等益生功能和作用机制的研究进展做一综述,并对研究前景提出展望,以期为我国乳酸菌的开发应用奠定基础。